Detecting antigen responsive cells in a sample

Abstract

The present invention relates to methods for detecting antigen responsive cells in a sample using multidimensional labeled antigen presenting compounds, such as antigen-major histocompatibility complexes (NHC). Further, the present invention relates to the use of the present multidimensional labeled antigen presenting compounds, such as antigen-major histocompatibility complexes (MHC), for detecting antigen responsive cells in a sample, preferably a single sample, such as a blood sample. The present method allows high-throughput analysis of specific antigen responsive cells, such as T- and B-cells, thereby providing, for example, high-throughput methods for monitoring of diseases or conditions and the development of immunotherapeutics, vaccines, or the identification epitopes or immunogenic amino acid sequences.

The invention claimed is: 
     
       1. A method for detecting antigen responsive cells in a sample comprising:
 providing antigen-containing antigen presenting compounds, each antigen-containing antigen presenting compound carrying at least one label, with two or more predetermined antigens, wherein each antigen is represented by at least two different labels; 
 contacting said antigen-containing antigen presenting compounds with said sample; 
 detecting binding of said antigen-containing antigen presenting compounds to said antigen responsive cells, thereby detecting cells responsive to at least one of said antigens; wherein the presence of the at least one of said antigens is detected by detecting a signal emitted by the at least two different labels bound to an antigen responsive cell through said antigen presenting compounds loaded with the at least one of said antigens. 
 
     
     
       2. The method according to  claim 1 , wherein said two or more predetermined antigens are selected from the group consisting of three or more, four or more, five or more, six or more, seven or more, eight or more, ten or more, eleven or more, twelve or more, thirteen or more, fourteen or more, fifteen or more, sixteen or more, seventeen or more, eighteen or more, nineteen or more, twenty or more, twenty or more, twenty-one or more, twenty-two or more, twenty-three or more, twenty-four or more, twenty-five or more, twenty-six or more, twenty-seven or more, and twenty-eight or more. 
     
     
       3. The method according to  claim 1 , wherein said antigen-containing antigen presenting compounds are provided with one label and at least one of said antigens is represented by at least two differently labelled antigen-containing antigen presenting compounds. 
     
     
       4. The method according to  claim 1 , wherein said antigen-containing antigen presenting compounds are provided with at least two different labels and at least one of said antigens is represented by one labelled antigen-containing antigen presenting compound. 
     
     
       5. The method according to  claim 1 , wherein at least one of said antigens is a peptide. 
     
     
       6. The method according to  claim 1 , wherein said antigen-containing antigen presenting compounds are major histocompatibility complexes (MHC). 
     
     
       7. The method according to  claim 6 , wherein said major histocompatibility complexes (MHC) are multimeric major histocompatibility complexes (MHC). 
     
     
       8. The method according to  claim 1 , wherein said antigen responsive cells are T-cells and/or B-cells. 
     
     
       9. The method according to  claim 1 , wherein said labels are fluorescent labels. 
     
     
       10. The method according to  claim 9 , wherein said fluorescent labels comprise qDots. 
     
     
       11. The method according to  claim 1 , wherein the number of different labels is selected from the group consisting three or more, four or more, five or more, six or more, seven or more, and eight or more. 
     
     
       12. The method according to  claim 1 , wherein at least one of said antigens is represented by at least three or at least four different labels. 
     
     
       13. The method according to  claim 1 , wherein said sample is a blood sample or a blood derived sample. 
     
     
       14. The method according to  claim 1 , wherein said detection comprises flow cytometry. 
     
     
       15. The method according to  claim 1 , wherein said at least two labels represent a single antigen for the detecting of antigen responsive cells in a sample. 
     
     
       16. The method according to  claim 1 , wherein at least one of said antigens comprises an epitope. 
     
     
       17. A method for detecting binding of a ligand to a cell or a cell-bound receptor comprising:
 labelling said ligand with at least one ligand-binding compound, the ligand-binding compound having at least one label, wherein the ligand is labelled with at least two different labels; and 
 detecting said cell or cell-bound receptor, wherein said cell or cell-bound receptor is detected by detecting a signal emitted by the at least two different labels based on binding of the cell or cell-bound receptor to said ligand.